127 results about "PHA binding" patented technology

The invention belongs to the technical field of biomedicine, and discloses an antinuclear antibody IgG indirect immunofluorescence detection method and system, and the detection standardization degree and the result reproducibility are significantly improved through the full-automatic system design in combination with precise mechanical transmission and an automatic sample adding device. Through an optimized FITC labeling system and a refined washing procedure, non-specific binding is remarkably reduced, and meanwhile, the binding efficiency of the antinuclear antibody and the secondary antibody is improved, so that the detection sensitivity is improved. Through the design of the automatic sample adding and sheet sealing device, the sample treatment efficiency is greatly improved, and simultaneous detection of large-scale samples can be realized. Through combination of a high-resolution imaging system and an intelligent image analysis algorithm, morphological characteristics and distribution rules of fluorescence signals are automatically extracted, and the accuracy and objectivity of a detection result are greatly improved.

The invention relates to the technical field of biological detection, and discloses a method for double-ring detection of lung cancer L858R mutation based on CRISPR / Cas9 and DNA, and the method comprises the following steps: enriching circulating tumor cells CTC from a blood sample; extracting genomic DNA (deoxyribonucleic acid) of the enriched CTC; carrying out amplification on the DNA fragment containing the L858R mutation by adopting recombinase polymerase amplification RPA (recombinase polymerase amplification); the amplified DNA fragment is hybridized with a recognition ring in a DNA double-ring structure, a DNA double ring is composed of the recognition ring and a report ring, and a part of region of the recognition ring is hybridized with a corresponding fragment of the report ring to form a staggered double-chain region; when L858R mutation exists, the mutated DNA and a recognition ring are hybridized to activate a CRISPR / Cas9 system, and under the guidance of sgRNA, Cas9 cuts hybrid double strands and releases a report ring; carrying out rolling circle amplification RCA under the action of DNA polymerase by taking the released report ring as a template; a fluorescent probe is used for detecting an RCA amplification product, and whether L858R mutation exists or not is judged according to a fluorescence signal. The CRISPR / Cas9 system is combined with a DNA double-ring structure, so that the ultra-sensitive and specific detection on the lung cancer L858R mutation is realized.

The present application relates to an anti-CD3 antibody and its application. The present application develops a specific antibody molecule targeting CD3 epsilon chain, which can activate T cells by combining with CD3 epsilon on T cells. A T cell binding protein containing the CD3 antibody is further designed, which can specifically target and bind to CD3 epsilon on the surface of T cells in vivo and in vitro, so as to achieve moderate activation of T cells. The CAR lentivirus containing the T cell binding protein of the present application can induce the generation of CAR-T cells in vitro and in vivo, and the CAR-T cells have good tumor cell killing efficiency.

The invention provides a method and a device for secondary filtration and re-removal of cholesterol ester after continuous blood purification and blood fat filtration, and belongs to the technical field of medical blood purification. According to the method and the device, the technology of combining enzymolysis, adsorption and membrane separation is adopted for residual CE in a CBP filtering product. The method comprises the following steps: firstly, decomposing CE into small molecules by using specific enzyme, and weakening the combination of CE and other components; residual impurities are further adsorbed through a high-performance adsorption material; and finally, efficient removal of CE is realized by virtue of a high-precision membrane separation technology, so that the residual quantity is reduced to be below a safety threshold value. Compared with a traditional technology, the problems that the CE-containing filtrate is high in recycling risk and low in resource utilization rate are effectively solved, the purity of a filtered product is remarkably improved, technical support is provided for industrial scenes such as plasma exchange, filtrate feedback, biopharmacy and extracorporeal circulation in the medical field, and the application prospect is wide. The safety and the resource utilization efficiency of the blood purification technology are promoted.

The invention relates to a method for splitting, assembling and site-specific integration of large-fragment exogenous DNA (deoxyribonucleic acid) in Trichoderma reesei. Comprising the following steps: (1) constructing an arg:: Ptcu-Cas9-TtrpC-hph strain containing a Cas9 protein; (2) constructing a pUC19 <-> integration site: a pyr4 plasmid; (3) obtaining a plurality of target gene segments containing integration site homologous arms and having overlapping regions at the tail ends of adjacent DNA segments; (4) obtaining a gRNA expression cassette of a targeted genome integration site; (5) arg conversion: the arg is converted into a Ptcu-Cas9-TtrpC-hph strain; and (6) screening and verifying. According to the method, the CRISPR / Cas9 system is combined with the TAR in the trichoderma reesei for the first time, the exogenous large-fragment DNA is split into a plurality of small fragments, then the plurality of small-fragment exogenous DNA are assembled in the trichoderma reesei and are subjected to fixed-point integration, one-step editing of the large-fragment exogenous DNA is realized, and compared with a traditional genetics method, the gene editing efficiency in the trichoderma reesei is greatly improved.

The invention discloses a one-step biosensor, and aims to solve the problems that a traditional detection device is tedious in steps and needs multiple operations to complete the detection process. The device mainly comprises a substrate layer and a channel layer, liquid with same or different specific biomolecules flows through multiple groups of gold particles through multiple groups of specific pipelines, so that the specific biomolecules are hung on the gold particles, whole blood in the channel layer is separated into plasma and blood cells through electrodes on the substrate layer, and after the plasma and a buffer solution are fully mixed, the blood cells in the channel layer are separated. The method comprises the following steps: enabling the plasma to flow through gold particles decorated by specific biomolecules, and combining target molecules in the plasma with the specific biomolecules to realize red shift and color change of SPR peaks, so that the same or different target molecules are identified, and the content of the target molecules is judged.

It is provided an anti-Aβ34 antibody and its use for diagnosing Alzheimer's disease in a patient, comprising obtaining a sample from the patient, detecting the level of Aβ34 in the sample by contacting the sample with the anti-Aβ34 antibody and detecting binding between Aβ34 and the antibody, and diagnosing the patient with Alzheimer's disease when the presence of Aβ34 in the sample is detected, alone or in combination with detecting an amyloid deposition marker such as Aβ42 and determining the ratio of Aβ34 / Aβ42.

The present application relates to the technical field of RNA extraction, and particularly relates to a pretreatment reagent, a method and a kit for RNA purification of whole blood samples. By introducing hydrophobic DESs as the core pretreatment component, the DESs are mixed with the whole blood sample in advance, the DESs form an independent phase layer after centrifugation by using the unique physicochemical properties, and the liposoluble drug components, inflammatory proteins and pigments are selectively extracted to the hydrophobic DESs phase and the RNA is preliminarily purified; the upper water phase of the pretreatment and purification is taken and added into a lysis solution containing a chaotropic salt to fully release the nucleic acid, and DNase I is introduced for digestion. The present application combines the pretreatment of hydrophobic DESs and the digestion of DNase I, and is used in combination with PVP (targeting adsorption of residual pigments and phenolic compounds), reduces the impurities of the lysis system through the front-end pretreatment, creates a low-interference and high-efficiency purification environment for the specific binding of RNA by the magnetic beads, and forms a synergistic removal network for the interference of complex samples.

The invention discloses an application of Siglec-10 / G or a fusion protein containing a Siglec-10 / G protein structure in preparation of a medicine for treating glioma. The application finds that Siglec-10 / G is a new key myeloid immune checkpoint molecule in a glioma immunosuppression microenvironment, and reveals a specific molecular mechanism that Siglec-10 / G and related action signals (such as MUC1, CKB and GOT1) thereof influence glioma generation. The Rebinant Mouse Siglec-10 (C-Fc) provided by the invention can be combined with a ligand on the surface of a tumor cell, so that endogenous Siglec-10 / G signal transduction on MDM is competitively blocked, the anti-tumor immunity of a tumor-bearing mouse is improved, the growth of glioma is inhibited, the survival time of the tumor-bearing mouse is prolonged, and the Rebinant Mouse Siglec-10 (C-Fc) can be used for treating glioma.

The application belongs to the technical field of biotechnology, and discloses a nano antibody targeting B cell BCMA and application thereof. The amino acid sequences of the complementarity determining regions CDR1-CDR3 of the nano antibody are selected from SEQ ID NO. 1-12. The nano antibody provided by the application can specifically bind to B cells in human blood, especially activated plasma cells, so as to eliminate B cells producing pathological antibodies. Compared with traditional antibodies, the nano antibody provided by the application has a smaller molecular weight, high stability, low cost, easy expression and high expression amount, and is suitable for large-scale production. The nano antibody is combined with a solid-phase carrier in the application, so that B cells in blood can be targeted and eliminated, the number of B cells in blood of patients can be reduced in a short time, and the antibody level can be reduced, thereby improving the quality of life, reducing complications, and providing a new treatment option for patients who are ineffective or intolerant to traditional treatment.

The invention belongs to the technical field of phosphatidylserine production, and particularly relates to immobilized phospholipase D based on biological imprinting and a preparation method and application thereof. The preparation method comprises the following steps: S1, dissolving L-serine in an acetic acid buffer solution to obtain a serine solution, adding a phospholipase D solution into the serine solution, and stirring to obtain a mixed solution; s2, adding sodium alginate into the mixed solution, stirring and dissolving to obtain a biopolymer solution; s3, adjusting the pH value of the chitosan solution to be neutral, then adding CaCl2 into the chitosan solution, and stirring and dissolving to obtain a CaCl2-containing chitosan solution; and S4, dropwise adding the biopolymer solution into a chitosan solution containing CaCl2, complexing to form microspheres, collecting the microspheres, and washing and drying the microspheres to obtain the immobilized phospholipase D based on biological imprinting. According to the method, the biological imprinting technology and the enzyme immobilization technology are combined, the immobilized PLD with high selectivity and high stability is prepared, and the phosphatidyl transfer reaction efficiency is improved.

The present application relates to a kind of sgRNA targeting Plagl1 gene DMR, epigenetic editing system and its application.Specifically, the present application provides sgRNA targeting Plagl1 gene differential methylation region, which forms epigenetic editing system with dCas9-Tet1-CD fusion protein.The system is loaded in injectable type light-cured silk fibroin hydrogel, the expression of Plagl1 gene in periosteum stem / progenitor cells is accurately activated by targeted demethylation modification, thereby promoting osteogenic differentiation.The present application combines epigenetic regulation with biomaterial delivery, and provides a safe, efficient new strategy for jaw bone defect regeneration and repair.

The invention discloses a solid-phase and liquid-phase combined synthesis method of tilpotide, and belongs to the field of chemical synthesis of polypeptide drugs. The method comprises the following steps: firstly, synthesizing a complete main peptide chain of the tiall peptide on a solid phase carrier, and protecting a side chain of lysine (Lys24) at the 24th site by adopting an orthogonal protecting group (Dde), so that side chain activated ester (Ecosanoic acid-gamma-Glu-AEA-AEA-OSu) can be specifically connected to an amino group of lysine (Lys20) at the 20th site, and the protecting group of the Lys24 can be selectively removed under a mild liquid phase condition to obtain crude peptide of the tiall peptide. Complex side chain modification is transferred from a solid phase to a liquid phase, the problems of steric hindrance and low connection efficiency are effectively solved, and the method has the advantages of being high in synthesis purity, few in side reaction, easy to amplify and the like and is suitable for industrial production.

The invention belongs to the technical field of biological medicines, and relates to a mutants of mu-conotoxin KIIIA rich in positive charges and application of the mutants. The mutant is obtained by implementing site-specific mutagenesis at the following positions on a mutant as shown in SEQ ID NO.1 in a sequence table: S6R; the amino acid sequence of the mutant is as shown in SEQ ID NO. 2 in a sequence table. According to the invention, systematic structural optimization is carried out on a parent peptide KIIIA containing three pairs of disulfide bonds, and a strategy of combining disulfide bond deletion and amino acid site-directed mutagenesis is adopted, so that a mutant which is simple and convenient to synthesize and has a remarkable inhibition effect on a NaV1.4 channel is obtained. The mutant has high activity, high stability and better pharmacokinetic characteristics, and has wide development potential.

Exemplary systems, methods, and computer-accessible medium are provided for Chimeric Antigen Receptor (CAR)-T therapy. Thus, the exemplary systems, methods, and computer-accessible medium are provided that select a first set of targets highly expressed in cancer and lowly in normal tissues, select a second set of targets that are highly co-expressed in the cancer and lowly in normal tissues, generate an input set by combining the first and second set of targets with dependency data, single-cell heterogeneity, and antibody availability, determine, by a learning model, at least one target pair for use in CAR-T therapy, and apply a cell therapy based on the determined target pair(s).

This invention belongs to the field of biomedical technology, specifically relating to a peptide targeting TRPV1 and its applications. A peptide targeting TRPV1 has the amino acid sequence shown in SEQ ID NO.1. The peptide also includes a mutant obtained by amino acid scanning mutation based on the sequence shown in SEQ ID NO.1. The amino acid sequence of the mutant is shown in any one of SEQ ID NO.2-16. This peptide is used to prepare formulations that block or inhibit TRPV1 activity. This invention provides a peptide targeting TRPV1 and its applications. Through a strategy combining artificial intelligence screening and structural prediction, SEQ ID NO.1 was successfully identified and its mechanism of action was elucidated. Experimental results show that SEQ ID NO.1 and its mutants exhibit excellent analgesic activity and good safety both in vitro and in vivo, demonstrating significant clinical application potential.

The application discloses a TYK2 inhibitor and a screening method and application thereof based on fusion of deep learning scoring and traditional molecular docking scoring. The application adopts a method of combining a deep learning scoring result CNNscore with an empirical scoring result obtained by Watvina, and improves the screening efficiency by multiplying the deep learning docking scoring and the empirical docking scoring, thereby relieving the problem of excessively high false positive rate of the traditional screening method, and saving time and cost. The application also applies the method to 12 million compounds, and verifies the compounds by in-vitro activity test, and obtains two small molecules with TYK2 inhibitory activity, which have application prospects in preventing or / and treating TYK2-mediated diseases. Therefore, the method is helpful to quickly identifying potential effective compounds from a huge compound library.

The invention belongs to the technical field of biological medicines, and relates to a mutants of mu-conotoxin KIIIA rich in positive charges and application of the mutants. The mutant is obtained by carrying out site-directed mutagenesis on C1A, K7R and C15R on a natural mu-conotoxin KIIIA sequence; the amino acid sequence of the mutant is shown as SEQ ID NO. 1 in a sequence table. According to the invention, a series of mutants which are simple and convenient to synthesize and have a remarkable inhibition effect on a NaV1.4 channel are obtained by performing systematic structural optimization on a parent peptide KIIIA containing three pairs of disulfide bonds and adopting a strategy of combining disulfide bond deletion and amino acid site-directed mutagenesis. Furthermore, non-natural amino acid Pen is introduced into the mutant with the optimal activity, so that the stability of the mutant in vivo is remarkably enhanced, and the action time is prolonged.

The invention discloses a human albumin binding peptide 4F1 and a method for purifying human albumin, and belongs to the technical field of polypeptides. The amino acid sequence of the human albumin binding peptide 4F1 comprises an amino acid sequence as shown in SEQ ID NO. 1; and / or an amino acid sequence of a fusion protein with the same function, which is obtained by connecting tag protein to the N terminal and / or C terminal of the amino acid sequence as shown in SEQ ID NO.1. The human albumin binding peptide 4F1 has extremely high affinity with human albumin and can be used for separating and purifying a human albumin solution, and the purity of the purified human albumin far exceeds the pharmacopoeia standard and can reach 99.9% or above. According to the method, high-selectivity capture of affinity chromatography and strong impurity removal capacity of gel filtration chromatography are combined, a purification process route which is efficient, stable and easy to amplify is constructed, and the recovery rate and the production efficiency are remarkably improved.

The present application relates to the technical field of biological medicines, and particularly relates to a polypeptide for specifically blocking the interaction between PD-L1 and endogenous double-stranded RNA and a medicine. The present application provides a polypeptide for specifically binding to endogenous double-stranded RNA, which has a strong effect on the activation of innate immune genes. The polypeptide fused with a cell-penetrating peptide can competitively bind to endogenous double-stranded RNA in cells and inhibit the A-to-I editing of endogenous double-stranded RNA, thereby improving the sensitivity of tumor cells to drugs. The present application provides a new strategy for the immunotherapy of tumor patients and has a wide application prospect.

The application provides a drug-target protein binding affinity prediction method and system, relates to the technical field of biological information technology, and comprises the following steps: learning local information of an atomic composite graph at a local level by using an isometric graph neural network; fully utilizing three-dimensional space structure information by using the isometric graph neural network, and keeping rotation and translation of coordinates unchanged during message transmission; then using a node feature extraction model and an atomic interaction matrix to learn deep-level interaction of the atomic composite graph at a global and long-distance level, improving interpretability of feature learning of the protein and the ligand, and making prediction performance of the drug-target protein binding affinity prediction model optimal. The application combines interaction of each distance level of the drug-target protein by fitting a dynamic process of drug-protein combination, simultaneously considers rotation and translation invariance of atomic mutual combination, and effectively improves accuracy of drug-target protein binding affinity prediction.

To provide a combination with previously-proven cancer therapies such as anti-angiogenic agents in order to improve clinical response rates to anti-PD1 antibodies.SOLUTION: An antibody or an antigen-binding fragment thereof that binds specifically to both PD1 and VEGFR2, and a pharmaceutical composition comprising the same, are provided.SELECTED DRAWING: None

A nanoagonist, and a preparation method and use thereof are provided, belonging to the technical field of nanoscale biomedicine. The nanoagonist is formed by self-assembly of a transformable peptide, where the transformable peptide includes a targeted antimicrobial peptide, a functionalized self-assembling peptide, an FcγR recognition peptide, and a lipase-responsive hydrophobic molecule that are coupled in sequence. The functionalized self-assembling peptide can control the FcγR recognition peptide to flip toward a surface of a target pathogen during secondary self-assembly, and the target pathogen is a pathogen targeted and bound by the targeted antimicrobial peptide. The nanoagonist combines externalization of the FcγR recognition peptide that can be guided during the secondary self-assembly with FcγR-mediated endocytosis, and a nanoagonist is developed for the first time that takes into account both pathogen clearance and host immune function repair.

Sotorasib augments the KRAS-G12C mutant cancer cells response to TNFα by Increasing TNFR1 surface expression, which in turn upregulates TNFα and IFNy down-stream target genes and ultimately enhances cancer cell death. In TACE-dependent mechanism, Sotorasib inhibited TNFR1 shedding off the KRAS-G12C mutant cancer cells. Additionally, Sotorasib significantly promotes the expansion of tumor infiltrating lymphocytes (TILs). In, addition, it generates a “hot” tumor microenvironment with strikingly augmented T cell effector phenotype. These results suggest that sotorasib regulation of TNFa and IFNy plays a crucial role in generating a more immune active tumor microenvironment. Consistently, sotorasib combined with CAR-T adoptive cell transfer and anti-PD1 treatment enhanced their anti-tumor effect. Therefore, disclosed herein is a method for treating KRAS-G12C mutant cancer in a subject, the method involving adoptively transferring an effective amount of autologous or allogeneic immune effector cells (T cells) to the subject in combination with a KRAS-G12C inhibitor

The present application provides a Morchella source hypoglycemic peptide and application thereof, and belongs to the technical field of biological medicine. The present application provides a Morchella source hypoglycemic peptide, which comprises a tripeptide and / or a tetrapeptide; the amino acid sequence of the tripeptide is PTW, and the amino acid sequence of the tetrapeptide is MPTW. The Morchella source hypoglycemic peptide of the present application has a molecular weight less than 1000, strong hydrophobicity, and contains Pro and Trp. Molecular docking results show that PTW and / or MPTW can be combined with the proteinase diabetes prevention and treatment target dipeptidyl peptidase IV, protein tyrosine phosphatase 1B and alpha-amylase to play a hypoglycemic role through hydrogen bonding and hydrophobic interaction. Moreover, the hypoglycemic peptide of the present application is a natural product, has no hemolyticity and allergenicity, and has good pharmacokinetic characteristics. The present application has certain guiding significance for the development of food source hypoglycemic peptides.

The application discloses the field of biotechnology, and discloses a kind of nanobody targeting B cell CD20 and application thereof.The amino acid sequence of nanobody complementarity determining region CDR1~CDR3 is selected from SEQ ID NO.1~9.The nanobody provided by the application can specifically bind to B cell in human blood, thereby eliminating B cell producing pathological antibody.The nanobody provided by the application has smaller molecular weight compared with traditional antibody, is stable, low in cost, easy to express and high in expression amount, and is suitable for large-scale production.Nanobody is combined with solid phase carrier in the application, B cell in blood can be targetedly eliminated, the number of B cell in blood of patient is reduced in a short time, antibody level is reduced, thereby improving life quality, reducing complication, and providing new treatment selection for those patients who are ineffective or intolerant to traditional treatment.

The invention discloses a construction method and application of a protein targeted degradation system, and belongs to the technical field of protein targeted degradation. According to the present invention, the Tris 21 Pturbo-GFP recombinant plasmid is constructed, and the Tris 21 Pturbo-GFP recombinant plasmid is combined with the electroporation delivery antibody technology so as to achieve the rapid degradation of the pig endogenous protein, and the dual plasmid co-transfection is combined with the electroporation delivery antibody to achieve the rapid degradation of the foreign protein; compared with traditional siRNA, the protein targeted degradation system has the advantages that the knock-down efficiency is improved by several times or more, and an excellent protein targeted degradation effect is achieved.